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Isolation and Culture of Single Myofiber and Immunostaining of Satellite Cells from Adult C57BL/6J Mice

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Intracellular Invasion and Killing Assay to Investigate the Effects of Binge Alcohol Toxicity in Murine Alveolar Macrophages

Authors: Victor Jimenez Jr and Fernando P Monroy, date: 01/20/2019, view: 998, Q&A: 0

[Abstract] Alcohol consumption has diverse and well-documented effects on the human immune system and its ability to defend against infective agents. While pulmonary related infections can occur in healthy humans, binge alcohol use is recognized as a major health risk factor (Nelson et al., 1991). Although binge alcohol consumption has been ...

Method for Studying the Effect of Gene Silencing on Bacterial Infection-induced ERK1/2 Signaling in Bone-marrow Derived Macrophages

Authors: Gaurav Kumar, Subhash B. Arya and Amit Tuli, date: 12/20/2018, view: 1386, Q&A: 0

[Abstract] Macrophages are highly phagocytic cells that utilize various pathogen recognition receptors (PRRs) to recognize pathogen-associated molecular patterns (PAMPs). These PAMPs can be present within the microbe, such as bacterial CpG DNA, and are recognized by Toll-like receptor 9 (TLR9), a PRR present on the endosomal membrane of macrophages. PAMPs ...

Phagocytosis Assay for α-Synuclein Fibril Uptake by Mouse Primary Microglia

Authors: Cuilian Du, Feifan Zhang and Claire Xi Zhang, date: 09/05/2018, view: 1341, Q&A: 0

[Abstract] Microglia are professional phagocytes in the brain and deficiency in their phagocytic activity plays an important role in Parkinson’s disease. This protocol mainly describes the phagocytosis assay for uptake of α-synuclein preformed fibrils, a pathologic form of α-synuclein, by primary microglia.

Measurement of TLR4 and CD14 Receptor Endocytosis Using Flow Cytometry

Authors: Michael S. Schappe and Bimal N. Desai, date: 07/20/2018, view: 2721, Q&A: 0

[Abstract] After recognizing extracellular bacterial lipopolysaccharide (LPS), the toll-like receptor 4 (TLR4)-CD14 signaling complex initiates two distinct signaling pathways–one from the plasma membrane and the other from the signaling endosomes (Kagan et al., 2008). Understanding the early stages of TLR4 signal transduction therefore requires a ...

3D Co-culture System of Tumor-associated Macrophages and Ovarian Cancer Cells

Authors: Lingli Long, Mingzhu Yin and Wang Min, date: 04/20/2018, view: 3333, Q&A: 0

[Abstract] Ovarian cancer is fairly unique in that ovarian carcinoma cells can detach and spread directly through peritoneal cavity. It has been unclear, however, how detached cancer cells survive in the peritoneum and form spheroid structure. We have recently reported that there is a strong correlation between Tumor-associated macrophages (TAMs)-associated ...

Flow Cytometric Quantification of Fatty Acid Uptake by Mycobacterium tuberculosis in Macrophages

Authors: Evgeniya V. Nazarova, Maria Podinovskaia, David G. Russell and Brian C. VanderVen, date: 02/20/2018, view: 3552, Q&A: 0

[Abstract] Mycobacterium tuberculosis (Mtb) has evolved to assimilate fatty acids from its host. However, until recently, there was no reliable way to quantify fatty acid uptake by the bacteria during host cell infection. Here we describe a new method to quantify fatty acid uptake by intracellular bacilli. We infect macrophages with Mtb ...

Phagocytosis Assay of Necroptotic Cells by Cardiac Myofibroblasts

Authors: Yuma Horii, Shoichi Matsuda, Kenji Watari, Akiomi Nagasaka, Hitoshi Kurose and Michio Nakaya, date: 09/20/2017, view: 3445, Q&A: 0

[Abstract] In myocardial infarction (MI), a plenty of cardiomyocytes undergo necrosis and necroptosis due to the lack of oxygen and nutrients. The dead cardiomyocytes are promptly engulfed by phagocytes. When the dead cells are not engulfed, the noxious contents of the cells are released outside, and thus, induce inflammation, and obstruct the function of ...

An Assay to Determine Phagocytosis of Apoptotic Cells by Cardiac Macrophages and Cardiac Myofibroblasts

Authors: Yuma Horii, Shoichi Matsuda, Kenji Watari, Akiomi Nagasaka, Hitoshi Kurose and Michio Nakaya, date: 09/20/2017, view: 3842, Q&A: 0

[Abstract] In myocardial infarction (MI), a number of cardiomyocytes undergo apoptosis. These apoptotic cardiomyocytes are promptly engulfed by phagocytes. If the dead cells are not engulfed, their noxious contents are released outside, resulting in induction of inflammation. Therefore, the removal of these dead cells is necessary. However, the contribution ...

Bioelectrospray Methodology for Dissection of the Host-pathogen Interaction in Human Tuberculosis

Authors: Liku B Tezera, Magdalena K Bielecka and Paul T Elkington, date: 07/20/2017, view: 4365, Q&A: 0

[Abstract] Standard cell culture models have been used to investigate disease pathology and to test new therapies for over fifty years. However, these model systems have often failed to mimic the changes occurring within three-dimensional (3-D) space where pathology occurs in vivo. To truthfully represent this, an emerging paradigm in biology is the ...

Creating a RAW264.7 CRISPR-Cas9 Genome Wide Library

Authors: Brooke A Napier and Denise M Monack, date: 05/20/2017, view: 6463, Q&A: 0

[Abstract] The bacterial clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 genome editing tools are used in mammalian cells to knock-out specific genes of interest to elucidate gene function. The CRISPR-Cas9 system requires that the mammalian cell expresses Cas9 endonuclease, guide RNA (gRNA) to lead the endonuclease to the gene of ...

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